Characterization of Recombinant Per a 10 from Periplaneta americana

Govindaraj, Dhanapal; Gaur, Shailendra Nath; Arora, Naveen

doi:10.1128/cvi.00461-12

Cited by 18 publications

(15 citation statements)

References 27 publications

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“…The trypsin-like sequences that were identified, as well as the sequence of the cloned cockroach trypsin (bgtryp-1) identified by Ock et al [12], were then used to deconvolute the sequences, based on the mass spectral analysis of the excised gel bands. Sequence alignment for the three germanica enzymes that we characterized, E1, E2 and E3, in comparison with each other and with the previously published cockroach sequences (bgtryp-1 and Per a 10 [8, 9, 12]) was done using the EMBL-EBI Clustal Omega online multiple sequence alignment tool that also generates the percent identity matrices (http://www.ebi.ac.uk/Tools/msa/clustalo/) [15]. The three B. germanica enzyme sequences were also compared in the same way for sequence alignment with the other Blattella antigen sequences in the database (Bla g 1 and Bla g 2) as well as the sequences of the dust mite proteinases, Der p 1, Der p 3 and Der p 6.…”

Section: Methodsmentioning

confidence: 99%

“…An important role for specific allergen proteinases was first appreciated when potent immunogenic antigens from the house dust mite ( Dermatophagoides pteronyssinus ), a major asthma risk-factor allergen, were found to have structural homology to cysteine (Der p 1) [3] and serine proteinases (Der p 3: tryptic Der p 6: chymotryptic and Der p 9: collagenolytic) [4]. A number of diverse environmental allergens have since been found to contain proteolytic activity, including several species of moulds [5], pollens [6], the German cockroach ( Blattella germanica ) [7], and the American cockroach ( Periplaneta americana ) [8, 9], among others. Previous work suggests that, similar to the house dust mite allergen, the proteinases in these other allergen sources may contribute to their allergenicity [10].…”

Section: Introductionmentioning

confidence: 99%

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Cockroach allergen serine proteinases: Isolation, sequencing and signalling via proteinase‐activated receptor‐2

Polley

Mihara

Ramachandran

et al. 2017

Clin Experimental Allergy

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SUMMARY BACKGROUND Allergy to the German cockroach (Blattella germanica) is a significant asthma risk factor for inner-city communities. Cockroach, like other allergens, contains trypsin-like enzyme activity that contributes to allergenicity and airway inflammation by activating proteinase-activated receptors (PARs). To date, the enzymes responsible for the proteolytic activity of German cockroach allergen have not been characterized. OBJECTIVES We aimed to identify, isolate and characterize the trypsin-like proteinases in German cockroach allergen extracts used for clinical skin tests. For each enzyme, we sought to determine (1) its substrate and inhibitor enzyme kinetics (Km and IC50); (2) its amino acid sequence and (3) its ability to activate calcium signaling and/or ERK1/2 phosphorylation via PAR2. METHODS Using a trypsin-specific activity-based probe, we detected three distinct enzymes that were isolated using ion-exchange chromatography. Each enzyme was sequenced by mass spectometery (deconvoluted with an expressed sequence tag library), evaluated kinetically for its substrate/inhibitor profile and assessed for its ability to activate PAR2 signaling. FINDINGS Each of the three serine proteinase-activity-based probe-labelled enzymes isolated were biochemically distinct, with different enzyme kinetic profiles and primary amino acid sequences. The three enzymes showed a 57 to 71% sequence identity with a proteinase previously cloned from the American cockroach (Per a 10). Each enzyme was found to activate both Ca++ and MAPK signaling via PAR2. CONCLUSIONS AND RELEVANCE We have identified three different serine proteinases from the German cockroach that may, via PAR2 activation, play different roles for allergen-sensitization in vivo and may represent attractive therapeutic targets for asthma.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Cockroach allergen serine proteinases: Isolation, sequencing and signalling via proteinase‐activated receptor‐2

Polley

Mihara

Ramachandran

et al. 2017

Clin Experimental Allergy

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“…Active Per a 10 lead to the secretion of proinflammatory cytokines from A549 cells in a dose and time dependent manner. Furthermore, inhibiting protease activity of Per a 10 by heat treatment or using a proteolytically inactive recombinant Per a 10 [11] failed to activate airway epithelial cells. To ascertain the role of protease activated receptors on secretion of proinflammatory cytokines, the epithelial cells were incubated with receptor cleavage blocking antibodies.…”

Section: Research Highlightmentioning

confidence: 98%

A cockroach derived serine protease allergen activates airway epithelial cells via PAR2 receptors

Kale

Arora

2014

Inflamm Cell Signal

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Both endogenous and exogenous proteases have been implicated in allergic airway diseases. Though recent studies have demonstrated the role of airway epithelial cells in pathophysiology of allergic diseases, the mechanisms of epithelial activation remain largely unknown. Recently published study (Kale and Arora, 2014) focused on elucidating the role of proteolytic activity of Per a 10 on airway epithelial activation and to gain an insight into possible mechanism. We found that airway epithelial activation by Per a 10 is dependent on its serine protease activity. Further we showed that this activation is PAR2 dependent and leads to Ca 2+ mobilisation. This Research Highlight discusses the findings of our recent study and active research endeavors.

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“…The Per a 10 protein sequence exhibited 27–38% similarity to mite serine proteases, and 41–52%similarity to other insect trypsins. The allergen showed potential for immunotherapy by having reduced IgE antibody binding and histamine release [84]. …”

Section: Cockroach Allergens Belong To Different Families Of Proteinsmentioning

confidence: 99%

Recombinant Allergens for Diagnosis of Cockroach Allergy

Arruda

Barbosa

Santos

et al. 2014

Curr Allergy Asthma Rep

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Molecular cloning of cockroach allergens and their expression as recombinant proteins have allowed a better understanding of the mechanisms of cockroach allergic disease. Recombinant cockroach allergens have been used for skin testing or in vitro methods to measure IgE antibody levels in serum. Early studies evaluating selected U.S. patients revealed that a cocktail of four cockroach allergens, Bla g 1, Bla g 2, Bla g 4, and Bla g 5, would identify 95 % of cockroach allergic patients. More recent studies pointed to an important role of sensitization to tropomyosin among certain populations, and suggested that a cocktail of five allergens Bla g 1 and/or Per a 1, Bla g 2, Bla g 4, Bla g 5, and Bla g 7, and/or Per a 7, would be expected to diagnose 50–64 % of cockroach-allergic patients worldwide. Variation in IgE reactivity profiles could be in part due to IgE responses to cross-reactive homologous allergens from different origins. The availability of purified natural or recombinant cockroach allergens provides the capacity to improve diagnosis of cockroach allergy and to develop novel forms of immunotherapy for cockroach-allergic patients.

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Characterization of Recombinant Per a 10 from Periplaneta americana

Cited by 18 publications

References 27 publications

Cockroach allergen serine proteinases: Isolation, sequencing and signalling via proteinase‐activated receptor‐2

Cockroach allergen serine proteinases: Isolation, sequencing and signalling via proteinase‐activated receptor‐2

A cockroach derived serine protease allergen activates airway epithelial cells via PAR2 receptors

Recombinant Allergens for Diagnosis of Cockroach Allergy

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